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Radical SAM enzymes / Edited by Vahe Bandarian
(Methods in enzymology ; v. 606)

First edition.
出版者 (Cambridge, MA : Academic Press)
出版年 [2018]
大きさ 1 online resource
書誌ID MC00035260

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eBook オンライン資料

MC000068784
Elsevier ScienceDirect Books Complete 9780128127957

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一般注記 Includes bibliographical references.
Online resource; title from PDF title page (EBSCO, viewed August 14, 2018)
Front Cover; Radical SAM Enzymes; Copyright; Contents; Contributors; Preface; Chapter One: Atlas of the Radical SAM Superfamily: Divergent Evolution of Function Using a ``Plug and Play" Domain; 1. Introduction: Overview of the Radical SAM Superfamily; 2. Results and Discussion; 2.1. Structural Overview; 2.2. A New Classification of the RSS; 2.3. A Large-Scale View of Sequence-Structure-Function Relationships; 2.4. Ancient Lineage of the RSS; 2.5. Classification of the RSS Based on Sequence Similarity versus Chemical Similarity; 2.6. Targeting Unknowns for Experimental Characterization
3. Methods3.1. Collection of RSS Sequences; 3.2. Representative Networks; 3.3. Determining Subgroups and Families; 3.4. Annotation of the RSS in the SFLD; Acknowledgments; Author Contributions; Competing Financial Interests Statement; References; Chapter Two: Purification and Characterization of the Choline Trimethylamine-Lyase (CutC)-Activating Protein CutD; 1. Introduction; 2. Overproduction and Purification of CutD; 2.1. Gene Cloning and Overexpression Strategy; 2.2. Protocol for Overexpression of the CutD Gene; 2.3. Protocol for Purifying CutD Under Anaerobic Conditions
2.4. Procedure for Chemical Reconstitution of Isolated CutD3. Characterization of the Iron-Sulfur Clusters in CutD; 3.1. UV-vis Spectrum of Reconstituted CutD; 3.2. Protocol for EPR Analysis of Iron-Sulfur Clusters; 3.3. Generation of SAM Cleavage Products Monitored by HPLC; 4. Activity of CutD Toward CutC; 4.1. Protocol for Quantification of Extent of CutC Activation by EPR; 4.2. Protocol for Kinetic Analysis of Choline Cleavage by CutC Upon Activation by CutD; 4.3. Protocol for LC-MS/MS-Based Detection of TMA From CutC-Catalyzed Choline Cleavage; 5. Conclusions; Acknowledgments; References
Chapter Three: QueE: A Radical SAM Enzyme Involved in the Biosynthesis of 7-Deazapurine Containing Natural Products1. Introduction; 2. Expression and Purification of QueE; 2.1. Expression of QueE; 2.2. Purification of QueE; 2.3. Chemical Reconstitution of QueE; 3. Expression and Purification of YkuN; 3.1. Expression of B. subtilis Flavodoxin YkuN; 3.2. Purification of YkuN; 4. Expression and Purification of FPR; 4.1. Expression of the fpr Gene; 4.2. Purification of FPR; 5. Enzymatic Synthesis of CPH4 With GCH I and QueD From GTP; 5.1. Expression and Purification of GCH I
5.2. Expression of QueD5.3. Purification of QueD; 5.4. Enzymatic Synthesis of CPH4; 6. Typical Assay Conditions for QueE; 6.1. Protocol for Assaying QueE In Vitro; 7. LC-MS Analysis of Turnover by CDG synthase; 8. Conclusions; Acknowledgments; References; Chapter Four: TYW1: A Radical SAM Enzyme Involved in the Biosynthesis of Wybutosine Bases; 1. Introduction; 2. Expression and Purification of TYW1; 2.1. Expression of TYW1 Gene; 2.1.1. Protocol for Expression of TYW1 Gene; 2.2. Purification of TYW1; 2.2.1. Protocol for Purification of His6-TYW1 Protein
Elsevier ScienceDirect Book Series Package - Methods in Enzymology (2000-ongoing)
著者標目 Bandarian, Vahe editor
件 名 LCSH:Adenosylmethionine
FREE:MEDICAL / Pharmacology
FREE:Adenosylmethionine
MESH:S-Adenosylmethionine
FREE:Electronic books
FREE:Electronic books
LCSH:Electronic books
分 類 LCC:QP563.A3
NLM:QU 57
UDC:577.15 Q8
DC23:615.78
資料種別 機械可読データファイル
巻冊次 ISBN:9780128127957
ISBN:0128127953
XISBN:9780128127940
XISBN:0128127945